The present invention relates to compositions comprising antibodies or fragments thereof that immunospecifically bind to a RSV antigen and methods for preventing, treating or ameliorating symptoms associated with respiratory syncytial virus (RSV) infection utilizing said compositions. In particular, the present invention relates to methods for preventing, treating or ameliorating symptoms associated with RSV infection, said methods comprising administering to a human subject an effective amount of one or more antibodies or fragments thereof that immunospecifically bind to a RSV antigen, wherein a certain serum titer of said antibodies or antibody fragments is achieved in said human subject. The present invention also relates to detectable or diagnostic compositions comprising antibodies or fragments thereof that immunospecifically bind to a RSV antigen and methods for detecting or diagnosing RSV infection utilizing said compositions.
Respiratory syncytial virus (RSV) is the leading cause of serious lower respiratory tract disease in infants and children (Feigen et al., eds., 1987, In: Textbook of Pediatric Infectious Diseases, WB Saunders, Philadelphia at pages 1653-1675; New Vaccine Development, Establishing Priorities, Vol. 1, 1985, National Academy Press, Washington D.C. at pages 397-409; and Ruuskanen et al., 1993, Curr. Probl. Pediatr. 23:50-79). The yearly epidemic nature of RSV infection is evident worldwide, but the incidence and severity of RSV disease in a given season vary by region (Hall, C. B., 1993, Contemp. Pediatr. 10:92-110). In temperate regions of the northern hemisphere, it usually begins in late fall and ends in late spring. Primary RSV infection occurs most often in children from 6 weeks to 2 years of age and uncommonly in the first 4 weeks of life during nosocomial epidemics (Hall et al., 1979, New Engl. J. Med. 300:393-396). Children at increased risk from RSV infection include preterm infants (Hall et al., 1979, New Engl. J. Med. 300:393-396) and children with bronchopulmonary dysplasia (Groothuis et al., 1988, Pediatrics 82:199-203), congenital heart disease (MacDonald et al., New Engl. J. Med. 307:397-400), congenital or acquired immunodeficiency (Ogra et al., 1988, Pediatr. Infect. Dis. J. 7:246-249; and Pohl et al., 1992, J. Infect. Dis. 165:166-169), and cystic fibrosis (Abman et al., 1988, J. Pediatr. 113:826-830). The fatality rate in infants with heart or lung disease who are hospitalized with RSV infection is 3%-4% (Navas et al., 1992, J. Pediatr. 121:348-354).
RSV infects adults as well as infants and children. In healthy adults, RSV causes predominantly upper respiratory tract disease. It has recently become evident that some adults, especially the elderly, have symptomatic RSV infections more frequently than had been previously reported (Evans, A. S., eds., 1989, Viral Infections of Humans. Epidemiology and Control, 3rd ed., Plenum Medical Book, New York at pages 525-544). Several epidemics also have been reported among nursing home patients and institutionalized young adults (Falsey, A. R., 1991, Infect. Control Hosp. Epidemiol. 12:602-608; and Garvie et al., 1980, Br. Med. J. 281:1253-1254). Finally, RSV may cause serious disease in immunosuppressed persons, particularly bone marrow transplant patients (Hertz et al., 1989, Medicine 68:269-281).
Treatment options for established RSV disease are limited. Severe RSV disease of the lower respiratory tract often requires considerable supportive care, including administration of humidified oxygen and respiratory assistance (Fields et al., eds, 1990, Fields Virology, 2nd ed., Vol. 1, Raven Press, New York at pages 1045-1072). The only drug approved for treatment of infection is the antiviral agent ribavirin (American Academy of Pediatrics Committee on Infectious Diseases, 1993, Pediatrics 92:501-504). It has been shown to be effective in the treatment of RSV pneumonia and bronchiolitis, modifying the course of severe RSV disease in immunocompetent children (Smith et al., 1991, New Engl. J. Med. 325:24-29). However, ribavirin has had limited use because it requires prolonged aerosol administration and because of concerns about its potential risk to pregnant women who may be exposed to the drug during its administration in hospital settings.
While a vaccine might prevent RSV infection, no vaccine is yet licensed for this indication. A major obstacle to vaccine development is safety. A formalin-inactivated vaccine, though immunogenic, unexpectedly caused a higher and more severe incidence of lower respiratory tract disease due to RSV in immunized infants than in infants immunized with a similarly prepared trivalent parainfluenza vaccine (Kim et al., 1969, Am. J. Epidemiol. 89:422-434; and Kapikian et al., 1969, Am. J. Epidemiol. 89:405-421). Several candidate RSV vaccines have been abandoned and others are under development (Murphy et al., 1994, Virus Res. 32:13-36), but even if safety issues are resolved, vaccine efficacy must also be improved. A number of problems remain to be solved. Immunization would be required in the immediate neonatal period since the peak incidence of lower respiratory tract disease occurs at 2-5 months of age. The immaturity of the neonatal immune response together with high titers of maternally acquired RSV antibody may be expected to reduce vaccine immunogenicity in the neonatal period (Murphy et al., 1988, J. Virol. 62:3907-3910; and Murphy et al., 1991, Vaccine 9:185-189). Finally, primary RSV infection and disease do not protect well against subsequent RSV disease (Henderson et al., 1979, New Engl. J. Med. 300:530-534).
Currently, the only approved approach to prophylaxis of RSV disease is passive immunization. Initial evidence suggesting a protective role for IgG was obtained from observations involving maternal antibody in ferrets (Prince, G. A., Ph.D. diss., University of California, Los Angeles, 1975) and humans (Lambrecht et al, 1976, J. Infect. Dis. 134:211-217; and Glezen et al., 1981, J. Pediatr. 98:708-715). Hemming et al. (Morell et al., eds., 1986, Clinical Use of Intravenous Immunoglobulins, Academic Press, London at pages 285-294) recognized the possible utility of RSV antibody in treatment or prevention of RSV infection during studies involving the pharmacokinetics of an intravenous immune globulin (IVIG) in newborns suspected of having neonatal sepsis. They noted that 1 infant, whose respiratory secretions yielded RSV, recovered rapidly after IVIG infusion. Subsequent analysis of the IVIG lot revealed an unusually high titer of RSV neutralizing antibody. This same group of investigators then examined the ability of hyperimmune serum or immune globulin, enriched for RSV neutralizing antibody, to protect cotton rats and primates against RSV infection (Prince et al., 1985, Virus Res. 3:193-206; Prince et al., 1990, J. Virol. 64:3091-3092; Hemming et al., 1985, J. Infect. Dis. 152:1083-1087; Prince et al., 1983, Infect. Immun. 42:81-87; and Prince et al., 1985, J. Virol. 55:517-520). Results of these studies suggested that RSV neutralizing antibody given prophylactically inhibited respiratory tract replication of RSV in cotton rats. When given therapeutically, RSV antibody reduced pulmonary viral replication both in cotton rats and in a nonhuman primate model. Furthermore, passive infusion of immune serum or immune globulin did not produce enhanced pulmonary pathology in cotton rats subsequently challenged with RSV.
Recent clinical studies have demonstrated the ability of this passively administered RSV hyperimmune globulin (RSV IVIG) to protect at-risk children from severe lower respiratory infection by RSV (Groothius et al., 1993, New Engl. J. Med. 329:1524-1530; and The PREVENT Study Group, 1997, Pediatrics 99:93-99). While this is a major advance in preventing RSV infection, this treatment poses certain limitations in its widespread use. First, RSV IVIG must be infused intravenously over several hours to achieve an effective dose. Second, the concentrations of active material in hyperimmune globulins are insufficient to treat adults at risk or most children with comprised cardiopulmonary function. Third, intravenous infusion necessitates monthly hospital visits during the RSV season. Finally, it may prove difficult to select sufficient donors to produce a hyperimmune globulin for RSV to meet the demand for this product. Currently, only approximately 8% of normal donors have RSV neutralizing antibody titers high enough to qualify for the production of hyperimmune globulin.
One way to improve the specific activity of the immunoglobulin would be to develop one or more highly potent RSV neutralizing monoclonal antibodies (MAbs). Such MAbs should be human or humanized in order to retain favorable pharmacokinetics and to avoid generating a human anti-mouse antibody response, as repeat dosing would be required throughout the RSV season. Two glycoproteins, F and G, on the surface of RSV have been shown to be targets of neutralizing antibodies (Fields et al., 1990, supra; and Murphy et al., 1994, supra). These two proteins are also primarily responsible for viral recognition and entry into target cells; G protein binds to a specific cellular receptor and the F protein promotes fusion of the virus with the cell. The F protein is also expressed on the surface of infected cells and is responsible for subsequent fusion with other cells leading to syncytia formation. Thus, antibodies to the F protein may directly neutralize virus or block entry of the virus into the cell or prevent syncytia formation. Although antigenic and structural differences between A and B subtypes have been described for both the G and F proteins, the more significant antigenic differences reside on the G glycoprotein, where amino acid sequences are only 53% homologous and antigenic relatedness is 5% (Walsh et al., 1987, J. Infect. Dis. 155:1198-1204; and Johnson et al., 1987, Proc. Natl. Acad. Sci. USA 84:5625-5629). Conversely, antibodies raised to the F protein show a high degree of cross-reactivity among subtype A and B viruses. Beeler and Coelingh (1989, J. Virol. 7:2941-2950) conducted an extensive analysis of 18 different murine MAbs directed to the RSV F protein. Comparison of the biologic and biochemical properties of these MAbs resulted in the identification of three distinct antigenic sites (designated A, B, and C). Neutralization studies were performed against a panel of RSV strains isolated from 1956 to 1985 that demonstrated that epitopes within antigenic sites A and C are highly conserved, while the epitopes of antigenic site B are variable.
A humanized antibody directed to an epitope in the A antigenic site of the F protein of RSV, SYNAGIS(copyright)palivizumab, is approved for intramuscular administration to pediatric patients for prevention of serious lower respiratory tract disease caused by RSV at recommended monthly doses of 15 mg/kg of body weight throughout the RSV season (November through April in the northern hemisphere). SYNAGIS(copyright) is a composite of human (95%) and murine (5%) antibody sequences. See, Johnson et al., 1997, J. Infect. Diseases 176:1215-1224 and U.S. Pat. No. 5,824,307, the entire contents of which are incorporated herein by reference. The human heavy chain sequence was derived from the constant domains of human IgG1 and the variable framework regions of the VH genes or Cor (Press et al., 1970, Biochem. J. 117:641-660) and Cess (Takashi et al., 1984, Proc. Natl. Acad. Sci. USA 81:194-198). The human light chain sequence was derived from the constant domain of Cxcexa and the variable framework regions of the VL gene K104 with Jxcexa-4 (Bentley et al., 1980, Nature 288:5194-5198). The murine sequences derived from a murine monoclonal antibody, Mab 1129 (Beeler et al., 1989, J. Virology 63:2941-2950), in a process which involved the grafting of the murine complementarity determining regions into the human antibody frameworks.
Although SYNAGIS(copyright) has been successfully used for the prevention of RSV infection in pediatric patients, multiple intramuscular doses of 15 mg/kg of SYNAGIS(trademark) is required to achieve a prophylactic effect. In pediatric patients less than 24 months of age, the mean half-life of SYNAGIS(copyright) has been shown to be 20 days and monthly intramuscular doses of 15 mg/kg have been shown to result in a mean xc2x1 standard derivation 30 day serum titer of 37xc2x121 xcexcg/ml after the first injection, 57xc2x141 xcexcg/ml after the second injection, 68xc2x151 xcexcg/ml after the third injection, and 72xc2x150 xcexcg/ml after the fourth injection (The IMpact RSV Study Group, 1998, Pediatrics 102:531-537). Serum concentrations of greater than 30 xcexcg/ml have been shown to be necessary to reduce pulmonary RSV replication by 100 fold in the cotton rat model of RSV infection. However, the administration of multiple intramuscular doses of 15 mg/kg of antibody is inconvenient for the patient. Thus, a need exists for antibodies that immunospecifically bind to a RSV antigen, which are highly potent, have an improved pharmacokinetic profile, and thus have an overall improved therapeutic profile. Further, a need exists for antibodies that immunospecifically bind to a RSV antigen which require less frequent administration.
Citation or discussion of a reference herein shall not be construed as an admission that such is prior art to the present invention.
The present invention is based, in part, on the development of methods for achieving or inducing a prophylactically or therapeutically effective serum titer of an antibody or fragment thereof that immunospecifically binds to a respiratory syncytial virus (RSV) antigen in a mammal by passive immunization with such an antibody or fragment thereof, which methods require lower dosages and/or less frequent administration than previously known methods. The present invention is also based, in part, on the identification of antibodies with higher affinities for a RSV antigen which result in increased efficacy for prophylactic or therapeutic uses such that lower serum titers are prophylactically or therapeutically effective, thereby permitting administration of lower dosages and/or reduced frequency of administration.
The present invention provides methods of preventing, neutralizing, treating and ameliorating one or more symptoms associated with RSV infection in a subject comprising administering to said subject one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with high affinity and/or high avidity. Because a lower serum titer of such antibodies or antibody fragments is therapeutically or prophylactically effective than the effective serum titer of known antibodies, lower doses of said antibodies or antibody fragments can be used to achieve a serum titer effective for the prevention, neutralization, treatment and the amelioration of symptoms associated with a RSV infection. The use of lower doses of antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens reduces the likelihood of adverse effects. Further, the high affinity and/or high avidity of the antibodies of the invention or fragments thereof enable less frequent administration of said antibodies or antibody fragments than previously thought to be necessary for the prevention, neutralization, treatment or the amelioration of symptoms associated with a RSV infection.
The present invention also provides antibodies which immunospecifically bind to one or more RSV antigens and have increased in vivo half-lives relative to known antibodies such as, e.g., SYNAGIS(copyright). In particular, the present invention encompasses antibodies which immunospecifically bind to one or more RSV antigens and have increased in vivo half-lives relative to known antibodies (e.g., SYNAGIS(copyright)), said increased half-lives resulting from one or more modifications (e.g., substitutions, deletions, or insertions) in amino acid residues identified to be involved in the interaction of the Fc domain of said antibodies and the FcRn receptor. The present invention also encompasses pegylated antibodies and fragments thereof which immunospecifically bind to one or more RSV antigens and have increased in vivo half-lives relative to known antibodies such as, e.g., SYNAGIS(copyright). The increased in vivo half-lives of antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens reduce the dosage and/or frequency of administration of said antibodies or fragments thereof to a subject.
The invention encompasses sustained release formulations for the administration of one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens to a subject. The sustained release formulations reduce the dosage and/or frequency of administration of said antibodies or antibody fragments to a subject. Further, the sustained release formulations may be administered to maintain a therapeutically or prophylactically effective serum titer which does not exceed a certain maximum serum titer for a certain period of time.
The present invention encompasses methods of delivering one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens directly to the site of RSV infection. In particular, the invention encompasses pulmonary delivery of one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens. The improved methods of delivering of one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens reduce the dosage and/or frequency of administration of said antibodies or antibody fragments to a subject.
The present invention provides antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have an association rate constant or kon rate (antibody (Ab)+antigen (Ag) Abxe2x88x92Ag) of at least 105 Mxe2x88x921sxe2x88x921, at least 5xc3x97105 Mxe2x88x921sxe2x88x921, at least 106 Mxe2x88x921sxe2x88x921, at least 5xc3x97106 Mxe2x88x921sxe2x88x921, at least 107 Mxe2x88x921sxe2x88x921, at least 5xc3x97107 Mxe2x88x921sxe2x88x921, or at least 108 Mxe2x88x921sxe2x88x921. In particular, the present invention provides compositions for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection, said compositions comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have an a kon rate of at least 105 Mxe2x88x921sxe2x88x921, at least 5xc3x97105 Mxe2x88x921sxe2x88x921, at least 106 Mxe2x88x921sxe2x88x921, at least 5xc3x97106 Mxe2x88x921sxe2x88x921, at least 107 Mxe2x88x921sxe2x88x921, at least 5xc3x97107 Mxe2x88x921sxe2x88x921, or at least 108 Mxe2x88x921sxe2x88x921.
The present invention provides antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have a koff rate (antibody (Ab)+antigen (Ag) Abxe2x88x92Ag) of less than 10xe2x88x921 sxe2x88x921, less than 5xc3x9710xe2x88x921 sxe2x88x921, less than 10xe2x88x922 sxe2x88x921, less than 5xc3x9710xe2x88x922 sxe2x88x921, less than 10xe2x88x923 sxe2x88x921, less than 5xc3x9710xe2x88x923 sxe2x88x921, less than 10xe2x88x924 sxe2x88x921, less than 5xc3x9710xe2x88x924 sxe2x88x921, less than 10xe2x88x925 sxe2x88x921, less than 5xc3x9710xe2x88x925 sxe2x88x921, less than 10xe2x88x926 sxe2x88x921, less than 5xc3x9710xe2x88x926 sxe2x88x921, less than 10xe2x88x927 sxe2x88x921, less than 5xc3x9710xe2x88x927 sxe2x88x921, less than 10xe2x88x928 sxe2x88x921, less than 5xc3x9710xe2x88x928 sxe2x88x921, less than 10xe2x88x929 sxe2x88x921, less than 5xc3x9710xe2x88x929 sxe2x88x921, or less than 10xe2x88x9210 sxe2x88x921. In particular, the present invention provides compositions for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection, said compositions comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have a koff rate of less than 10xe2x88x921 sxe2x88x921, less than 5xc3x9710xe2x88x921 sxe2x88x921, less than 10xe2x88x922 sxe2x88x921, less than 5xc3x9710xe2x88x922 sxe2x88x921, less than 10xe2x88x923 sxe2x88x921, less than 5xc3x9710xe2x88x923 sxe2x88x921, less than 10xe2x88x924 sxe2x88x921, less than 5xc3x9710xe2x88x924 sxe2x88x921, less than 10xe2x88x925 sxe2x88x921, less than 5xc3x9710xe2x88x925 sxe2x88x921, less than 10xe2x88x926 sxe2x88x921, less than 5xc3x9710xe2x88x926 sxe2x88x921, less than 10xe2x88x927 sxe2x88x921, less than 5xc3x9710xe2x88x927 sxe2x88x921, less than 10xe2x88x928 sxe2x88x921, less than 5xc3x9710xe2x88x928 sxe2x88x921, less than 10xe2x88x929 sxe2x88x921, less than 5xc3x9710xe2x88x929 sxe2x88x921, or less than 10xe2x88x9210 sxe2x88x921.
The present invention also provides antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have an affinity constant or Ka (kon/koff) of at least 102 Mxe2x88x921, at least 5xc3x97102 Mxe2x88x921, at least 103 Mxe2x88x921, at least 5xc3x97103 Mxe2x88x921, at least 104 Mxe2x88x921, at least 5xc3x97104 Mxe2x88x921, at least 105 Mxe2x88x921, at least 5xc3x97105 Mxe2x88x921, at least 106 Mxe2x88x921, at least 5xc3x97106 Mxe2x88x921, at least 107 Mxe2x88x921, at least 5xc3x97107 Mxe2x88x921, at least 108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, at least 5xc3x971012 Mxe2x88x921, at least 1013 Mxe2x88x921, at least 5xc3x971013 Mxe2x88x921, at least 1014 Mxe2x88x921, at least 5xc3x971014 Mxe2x88x921, at least 1015 Mxe2x88x921, or at least 5xc3x971015 Mxe2x88x921. In particular, the present invention provides compositions for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection, said compositions comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have a Ka of at least 102 Mxe2x88x921, at least 5xc3x97102 Mxe2x88x921, at least 103 Mxe2x88x921, at least 5xc3x97103 Mxe2x88x921, at least 104 Mxe2x88x921, at least 5xc3x97104 Mxe2x88x921, at least 105 Mxe2x88x921, at least 5xc3x97105 Mxe2x88x921, at least 106 Mxe2x88x921, at least 5xc3x97106 Mxe2x88x921, at least 107 Mxe2x88x921, at least 5xc3x97107 Mxe2x88x921, at least 108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, at least 5xc3x971012 Mxe2x88x921, at least 1013 Mxe2x88x921, at least 5xc3x971013 Mxe2x88x921, at least 1014 Mxe2x88x921, at least 5xc3x971014 Mxe2x88x921, at least 1015 Mxe2x88x921, or at least 5xc3x971015 Mxe2x88x921.
The present invention provides antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have a median effective concentration (EC50) of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay. In particular, the present invention provides compositions for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection, said compositions comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and have an EC50 of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay
The present invention also provides antibodies or fragments thereof comprising a VH domain having the amino acid sequence of any VH domain listed in Table 2 and compositions comprising said antibodies or antibody fragments for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection. The present invention also provides antibodies or fragments thereof comprising one or more VH complementarity determining regions (CDRs) having the amino acid sequence of one or more VH CDRs listed in Table 2 and/or Table 3 and compositions comprising said antibodies or antibody fragments for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection. The present invention also provides antibodies or fragments thereof comprising a VL domain having the amino acid sequence of any VL domain listed in Table 2. The present invention also provides antibodies or fragments thereof comprising one or more VL CDRs having the amino acid sequence of one or more VL CDRs listed in Table 2 and/or Table 3 and compositions comprising said antibodies or antibody fragments for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection. The present invention further provides antibodies comprising a VH domain and a VL domain having the amino acid sequence of any VH domain and VL domain listed in Table 2 and compositions comprising said antibodies or antibody fragments for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection. The present invention further provides antibodies comprising one or more VH CDRs and one or more VL CDRs having the amino acid sequence of one or more VH CDRs and one or more VL CDRs listed in Table 2 and/or 3 and compositions comprising said antibodies or antibody fragments for use in the prevention, treatment or amelioration of one or more symptoms associated with a RSV infection. In the above embodiments, preferably the antibody binds immunospecifically to a RSV antigen.
The present invention also encompasses methods for achieving a serum titer of at least 40 xcexcg/ml of one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens in a mammal, preferably a primate and most preferably a human. In particular, the present invention provides methods for achieving a serum titer of at least 40 xcexcg/ml (preferably at least 75 xcexcg/ml, more preferably at least 100 xcexcg/ml, and most preferably at least 150 xcexcg/ml) of an antibody or fragment thereof that immunospecifically binds to a RSV antigen in a non-primate mammal, comprising administering a dose of less than 2.5 mg/kg (preferably 1.5 mg/kg or less) of the antibody or antibody fragment to the non-primate mammal and measuring the serum titer of the antibody or antibody fragment at least 1 day after administering the dose to the non-primate mammal. The present invention also provides methods for achieving a serum titer of at least 150 xcexcg/ml (preferably at least 200 xcexcg/ml) of an antibody or fragment thereof that immunospecifically binds to a RSV antigen in a non-primate mammal, comprising administering a dose of approximately 5 mg/kg of the antibody or antibody fragment to the non-primate mammal and measuring the serum titer of the antibody or antibody fragment at least 1 day after the administration of the dose to the non-primate mammal.
The present invention also provides methods for achieving a serum titer of at least 40 xcexcg/ml of an antibody or fragment thereof that immunospecifically binds to a RSV antigen in a primate, comprising administering a first dose of 10 mg/kg (preferably 5 mg/kg or less and more preferably 1.5 mg/kg or less) of the antibody or antibody fragment to the primate and measuring the serum titer of the antibody or antibody fragment 20 days (preferably 25, 30, 35 or 40 days) after administrating the first dose to the primate and prior to the administration of any subsequent dose. The present invention also provides methods for achieving a serum titer of at least 75 xcexcg/ml (preferably at least 100 xcexcg/ml, at least 150 xcexcg/ml, or at least 200 xcexcg/ml) of an antibody or fragment thereof that immunospecifically binds to a RSV antigen in a primate, comprising administering a first dose of approximately 15 mg/kg of the antibody or antibody fragment to the primate and measuring the serum titer of the antibody or antibody fragment 20 days (preferably 25, 30, 35 or 40 days) after administering the first dose to the primate but prior to any subsequent dose.
The present invention also provides methods for preventing, treating, or ameliorating one or more symptoms associated with a RSV infection in a human subject, said methods comprising administering to said human subject at least a first dose of approximately 15 mg/kg of an antibody or fragment thereof that immunospecifically binds to a RSV antigen so that said human subject has a serum antibody titer of at least 75 xcexcg/ml, preferably at least 100 xcexcg/ml, at least 150 xcexcg/ml, or at least 200 xcexcg/ml 30 days after the administration of the first dose of the antibody or antibody fragment and prior to the administration of a subsequent dose. The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a human subject, said methods comprising administering to said human subject at least a first dose of less than 15 mg/kg (preferably 10 mg/kg or less, more preferably 5 mg/kg or less, and most preferably 1.5 mg/kg or less) of an antibody or fragment thereof that immunospecifically binds to a RSV antigen so that said human subject has a serum antibody titer of at least 75 xcexcg/ml, preferably at least 100 xcexcg/ml, at least 150 xcexcg/ml, or at least 200 xcexcg/ml 30 days after the administration of the first dose of the antibody or antibody fragment and prior to the administration of a subsequent dose. The present invention further provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a human subject, said methods comprising administering to said human subject a first dose of an antibody or fragment thereof that immunospecifically binds to a RSV antigen such that a prophylactically or therapeutically effective serum titer of less than 10 xcexcg/ml is achieved no more than 30 days after administering the antibody or antibody fragment.
The present invention provides methods for achieving a therapeutically or prophylactically effective serum titer in a mammal, said methods comprising administering to said mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has a kon rate (antibody (Ab)+antigen (Ag) Abxe2x88x92Ag) of at least 2.5xc3x9710xe2x88x925Mxe2x88x921 s1, preferably at least 3xc3x97105Mxe2x88x921 sxe2x88x921, at least 5xc3x97105Mxe2x88x921 sxe2x88x921, at least 106Mxe2x88x921 sxe2x88x921, at least 5xc3x97106Mxe2x88x921 sxe2x88x921, at least 107Mxe2x88x921 sxe2x88x921, at least 5xc3x97107Mxe2x88x921 sxe2x88x921 or at least 108Mxe2x88x921 sxe2x88x921. In particular, the present invention provides methods for achieving a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period, comprising administering to a mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has a kon rate of at least 2.5xc3x97105 Mxe2x88x921sxe2x88x921, preferably at least 3xc3x97105 Mxe2x88x921sxe2x88x921, at least 5xc3x97105 Mxe2x88x921sxe2x88x921, at least 106 Mxe2x88x921sxe2x88x921, at least 5xc3x97106 Mxe2x88x921sxe2x88x921, at least 107 Mxe2x88x921sxe2x88x921, at least 5xc3x97107 Mxe2x88x921sxe2x88x921, or at least 108 Mxe2x88x921sxe2x88x921. Preferably, the antibody or antibody fragment has a higher kon rate than SYNAGIS(copyright).
The present invention also provides methods of neutralizing RSV using an antibody or fragment thereof which immunospecifically bind to a RSV antigen and which has a kon rate of at least 2.5xc3x97105 Mxe2x88x921sxe2x88x921, preferably at least 3xc3x97105 Mxe2x88x921sxe2x88x921, at least 5xc3x97105 Mxe2x88x921sxe2x88x921, at least 106 Mxe2x88x921sxe2x88x921, at least 5xc3x97106 Mxe2x88x921sxe2x88x921, at least 107 Mxe2x88x921sxe2x88x921, at least 5xc3x97107 Mxe2x88x921sxe2x88x921, or at least 108 Mxe2x88x921sxe2x88x921 to achieve a prophylactically or therapeutically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) 20, 25, 30, or 35 days after administration without any other dosage administration. Preferably, the antibody or antibody fragment has a higher kon rate than SYNAGIS(copyright).
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, said methods comprising administering to said mammal, a dose of less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of an antibody or fragment thereof which immunospecifically binds to a RSV antigen and has a kon rate of at least 2.5xc3x97105 Mxe2x88x921sxe2x88x921, preferably at least 3xc3x97105 Mxe2x88x921sxe2x88x921, at least 5xc3x97105 Mxe2x88x921sxe2x88x921, at least 106 Mxe2x88x921sxe2x88x921, at least 5xc3x97106 Mxe2x88x921sxe2x88x921, at least 107 Mxe2x88x921sxe2x88x921, at least 5xc3x97107 Mxe2x88x921sxe2x88x921, or at least 108 Mxe2x88x921sxe2x88x921. Preferably, the antibody or antibody fragment has a higher kon rate for the RSV F glycoprotein than SYNAGIS(copyright).
The present invention also provides methods for achieving a therapeutically or prophylactically effective serum titer in a mammal, said methods comprising administering to said mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has a Koff rate (antibody (Ab)+antigen (Ag) Abxe2x88x92Ag) of less than 6.5xc3x9710xe2x88x924 secxe2x88x921, less than 5xc3x9710xe2x88x924 secxe2x88x921, less than 3xc3x9710xe2x88x924 secxe2x88x921, less than 2xc3x9710xe2x88x924 secxe2x88x921, less than 1xc3x9710xe2x88x924 secxe2x88x921, or less than 5xc3x9710xe2x88x923 secxe2x88x921. In particular, the present invention provides methods for achieving a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period, comprising administering to a mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has a Koff rate of less than 6.5xc3x9710xe2x88x924 secxe2x88x921, less than 5xc3x9710xe2x88x924 secxe2x88x921, less than 3xc3x9710xe2x88x924 secxe2x88x921, less than 2xc3x9710xe2x88x924 secxe2x88x921, less than 1xc3x9710xe2x88x924 secxe2x88x921, or less than 3xc3x9710xe2x88x923 secxe2x88x921. Preferably, the antibody or fragment thereof has a lower Koff rate than SYNAGIS(copyright).
The present invention also provides methods of neutralizing RSV using an antibody or antibody fragment thereof which immunospecifically binds to a RSV antigen and which has a Koff rate of less than 6.5xc3x9710xe2x88x924 secxe2x88x921, less than 5xc3x9710xe2x88x924 secxe2x88x921, less than 3xc3x9710xe2x88x924 secxe2x88x921, less than 2xc3x9710xe2x88x924 secxe2x88x921, less than 1xc3x9710xe2x88x924 secxe2x88x921, or less than 5xc3x9710xe2x88x923 secxe2x88x921 to achieve a prophylactically or therapeutically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) 20, 25, 30, or 35 days after administration without any other dosage administration. Preferably, the antibody or antibody fragment has a lower Koff than SYNAGIS(copyright).
The present invention also provides methods for preventing, treating, or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, said methods comprising administering to a said mammal a dose of less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of an antibody or a fragment thereof which immunospecifically binds to a RSV antigen and which has a Koff rate of less than 6.5xc3x9710xe2x88x924 secxe2x88x921, less than 5xc3x9710xe2x88x924 secxe2x88x921, less than 3xc3x9710xe2x88x924 secxe2x88x921, less than 2xc3x9710xe2x88x924 secxe2x88x921, less than 1xc3x9710xe2x88x924 secxe2x88x921, or less than 5xc3x9710xe2x88x923 secxe2x88x921. Preferably, the antibody or antibody fragment has a lower Koff rate than SYNAGIS(copyright).
The present invention also provides methods for achieving a therapeutically or prophylactically effective serum titer in a mammal, said methods comprising administering to said mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has an EC50 of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay. In particular, the present invention provides methods for achieving a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period, comprising administering to a mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has an EC50 of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay Preferably, the antibody or antibody fragment has a lower EC50 than SYNAGIS(copyright).
The present invention also provides methods of neutralizing RSV using an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has an EC50 of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay to achieve a prophylactically or therapeutically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) 20, 25, 30, or 35 days after administration without any other dosage administration. Preferably, the antibody or antibody fragment has a lower EC50 than SYNAGIS(copyright).
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, said methods comprising administering to said mammal a dose of less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of an antibody or a fragment thereof which immunospecifically binds to a RSV antigen and which has an EC50 of less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitro microneutralization assay. Preferably, the antibody or antibody fragment has a lower EC50 than SYNAGIS(copyright).
The present invention provides methods for achieving a therapeutically or prophylactically effective serum titer in a mammal, said methods comprising administering to said mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen and which has an affinity constant (Ka) for a RSV antigen of at least 2xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, at least 5xc3x971012 Mxe2x88x921, at least 1013 Mxe2x88x921, at least 5xc3x971013 Mxe2x88x921, at least 1014 Mxe2x88x921, at least 5xc3x971014 Mxe2x88x921, at least 1015 Mxe2x88x921, or at least 5xc3x971015 Mxe2x88x921. In particular, the present invention also provides methods for achieving a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period, comprising administering to a mammal an antibody or fragment thereof that has an affinity constant (Ka) for a RSV antigen of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, at least 5xc3x971012 Mxe2x88x921, at least 1013 Mxe2x88x921, at least 5xc3x971013 Mxe2x88x921, at least 1014 Mxe2x88x921, at least 5xc3x971014 Mxe2x88x921, at least 1015 Mxe2x88x921, or at least 5xc3x971015 Mxe2x88x921. Preferably, the antibody or antibody fragment has a higher affinity for a RSV F glycoprotein than SYNAGIS(copyright).
The present invention also provides methods of achieving a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period, comprising administering to a mammal an antibody or fragment thereof which immunospecifically binds to a RSV antigen with a higher avidity than known antibodies such as, e.g., SYNAGIS(copyright).
The present invention also provides methods of neutralizing RSV using an antibody or fragment thereof that has an affinity constant (Ka) for a RSV antigen of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, at least 5xc3x971012 Mxe2x88x921, at least 1013 Mxe2x88x921, at least 5xc3x971013 Mxe2x88x921, at least 1014 Mxe2x88x921, at least 5xc3x971014 Mxe2x88x921, at least 1015 Mxe2x88x921, or at least 5xc3x971015 Mxe2x88x921 to achieve a prophylactically or therapeutically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is at least 2 xcexcg/ml and more preferably at least 6 xcexcg/ml) 20, 25, 30, or 35 days after administration without any other dosage administration. Preferably, the antibody or antibody fragment has a higher affinity for the RSV F glycoprotein than SYNAGIS(copyright). The present invention also provides methods of neutralizing RSV using an antibody or fragment thereof that has a higher avidity than known antibodies such as, e.g., SYNAGIS(copyright).
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, said methods comprising administering to said mammal a dose of less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of an antibody or fragment thereof that has an affinity constant (Ka) for a RSV antigen of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, at least 5xc3x971012 Mxe2x88x921, at least 1013 Mxe2x88x921, at least 5xc3x971013 Mxe2x88x921, at least 1014 Mxe2x88x921, at least 5xc3x971014 Mxe2x88x921, at least 1015 Mxe2x88x921, or at least 5xc3x971015 Mxe2x88x921. Preferably, the antibody or antibody fragment has a higher affinity for the RSV F glycoprotein than SYNAGIS(copyright). The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, said methods comprising administering to said mammal a first dose of less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of an antibody or fragment thereof that has a higher avidity than known antibodies such as, e.g., SYNAGIS(copyright).
The present invention encompasses methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, comprising administering to said mammal a first dose of a prophylactically or therapeutically effective amount of one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright), wherein said effective amount is less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of said antibodies or antibody fragments which dose results in a serum titer of less than 30 xcexcg/ml (which is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) at least 20 days (preferably at least 25, at least 30, or at least 35 days) after the administration of the first dose and prior to the administration of a subsequent dose. In particular, the present invention provides methods for preventing, treating, or ameliorating one or more symptoms associated with a RSV infection in a human subject, comprising administering to said human subject a first dose of less than 5 mg/kg (preferably 3 mg/kg or less, and most preferably 1.5 mg/kg) of an antibody or fragment thereof that immunospecifically binds to a RSV antigen with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921) so that said human subject has a serum antibody titer of less than 30 xcexcg/ml (which is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) at least 20 days (preferably at least 25, at least 30, or at least 35 days) after the administration of the first dose of the antibody or antibody fragment and prior to the administration of a subsequent dose.
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising a VH domain having an amino acid sequence of any VH domain listed in Table 2 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising one or more VH complementarity determining regions (CDRs) having the amino acid sequence of one or more VH CDRs listed in Table 2 and/or Table 3 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. Preferably, said antibodies or antibody fragments immunospecifically bind to a RSV antigen.
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising a VL domain having the amino acid sequence of any VL domain listed in Table 2 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising one or more VL CDRs having the amino acid sequence of one or more VL CDRs listed in Table 2 and/or Table 3 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. Preferably, said antibodies or antibody fragments immunospecifically bind to a RSV antigen.
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising a VH domain and a VL domain having the amino acid sequence of any VH domain and VL domain listed in Table 2 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising one or more VH CDRs and one or more VL CDRs having the amino acid sequence of one or more VH CDRs and one or more VL CDRs listed in Table 2 and/or 3 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. Preferably, said antibodies or antibody fragments immunospecifically bind to a RSV antigen.
In a specific embodiment, the present invention provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising a VH domain having an amino acid sequence of SEQ ID NO:7, 9, 17, 24, 28, 33, 36, 40, 44, 48, 51, 67, or 78 and/or a VL domain having an amino acid sequence of SEQ ID NO:8, 11, 13, 21, 26, 30, 34, 38, 42, 46, 49, 52, 54, 56, 58, 60, 62, 64, 65, 68, 70, 71, 74 or 76 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. In a preferred embodiment, the present invention provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising a VH domain having an amino acid sequence of SEQ ID NO:9, 17, 24, 28, 33, 36, 40, 44, 48, 51, 55, 67 or 78 and/or a VL domain having an amino acid sequence of SEQ ID NO:13, 21, 26, 30, 34, 38, 42, 46, 49, 52, 54, 56, 58, 60, 62, 64, 65, 68, 70, 71, 74 or 76 to achieve a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period. In another embodiment, the present invention provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, said methods comprising administering to said mammal a first dose of one or more antibodies or fragments thereof comprising a VH CDR3 having an amino acid sequence of SEQ ID NO:3, 12, 20, 29, or 79 and a VL CDR3 having an amino acid sequence of SEQ ID NO:6, 16 or 61 to a therapeutically or prophylactically effective serum titer, wherein said effective serum titer is less than 30 xcexcg/ml (and is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) after a certain number of days (for example, but not limited to, 20, 25, 30 or 35 days) without any other dosing within that period.
The present invention also provides compositions comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and which have increased in vivo half-lives compared to known anti-RSV antibodies as a result of, e.g., one or more modifications in amino acid residues identified to be involved in the interaction between the Fc domain of said antibodies or antibody fragments and the FcRn receptor. In one embodiment, a composition of the invention comprises HL-SYNAGIS or an antigen-binding fragment thereof. In another embodiment, a composition of the invention comprises one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with a higher avidity and/or a higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) and which comprise an Fc domain with increased affinity for the FcRn receptor relative to the Fc domain of SYNAGIS(copyright). In accordance with this embodiment, the increased affinity of the Fc domain of said antibodies or antibody fragments results in an in vivo half-life of said antibodies or antibody fragments of at least 25 days, preferably at least 30 days, more preferably at least 30 days, and most preferably at least 40 days. In another embodiment, a composition of the invention comprises HL-SYNAGIS or an antigen-binding fragment thereof and one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and which comprise an Fc domain with increased affinity for the FcRn receptor relative to the Fc domain of SYNAGIS(copyright).
The present invention also provides compositions comprising one or more pegylated antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens. In one embodiment, a composition of the invention comprises pegylated SYNAGIS(copyright) or a fragment thereof. In another embodiment, a composition of the invention comprises one or more pegylated antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright). In yet another embodiment, a composition of the invention comprises pegylated SYNAGIS(copyright) or an antigen-binding fragment thereof and one or more pegylated antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright).
The present invention also provides compositions comprising one or more pegylated antibodies or fragments thereof which comprise an Fc domain with increased affinity for the FcRn receptor relative to the Fc domain of SYNAGIS(copyright). In one embodiment, a composition of the invention comprises a pegylated HL-SYNAGIS or an antigen-binding fragment thereof. In another embodiment, a composition of the invention comprises one or more pegylated antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with a higher avidity and/or a higher affinity than known such as, e.g., SYNAGIS(copyright) and which comprise an Fc domain with increased affinity for the FcRn receptor relative to the Fc domain of SYNAGIS(copyright).
The present invention encompasses methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, comprising administering to said mammal a first dose of a prophylactically or therapeutically effective amount of HL-SYNAGIS or an antigen-binding fragment thereof, wherein said effective amount is approximately 15 mg/kg of said antibodies or fragments thereof which dose results in a serum titer of at least 30 xcexcg/ml at least 30 days after the administration of the first dose and prior to the administration of a subsequent dose. In particular, the present invention provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a human subject, comprising administering to said human subject a first dose of 15 mg/kg of HL-SYNAGIS or an antigen-binding fragment thereof so that said human subject has a serum antibody titer of at least 30 xcexcg/ml at least 30 days after the administration of the first dose of the antibody or antibody fragment and prior to the administration of a subsequent dose.
The present invention also encompasses methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, comprising administering to said mammal a first dose of a prophylactically or therapeutically effective amount of one or more antibodies or fragments thereof which have increased in vivo half-lives and which immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen), wherein said effective amount is less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of said antibodies or fragments thereof which dose results in a serum titer of less than 30 xcexcg/ml (which is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) at least 20 days (preferably at least 25, at least 30, or at least 35 days) after the administration of the first dose and prior to the administration of a subsequent dose. In particular, the present invention provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a human subject, comprising administering to said human subject a first dose of less than 5 mg/kg (preferably 1.5 mg/kg or less) of an antibody or a fragment thereof which has an increased in vivo half-life and which immunospecifically binds to a RSV antigen with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) so that said human subject has a serum antibody titer of less than 30 xcexcg/ml (which is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) at least 25 days (preferably at least 30, at least 35, or at least 40 days) after the administration of the first dose of the antibody or antibody fragment and prior to the administration of a subsequent dose.
The present invention provides sustained release formulations comprising one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens. In one embodiment, a sustained release formulation comprises SYNAGIS(copyright) or a fragment thereof. In another embodiment, a sustained release formulation comprises one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen). In another embodiment, a sustained release formulation comprises SYNAGIS(copyright) or an antigen-binding fragment thereof and one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen). In another embodiment, HL-SYNAGIS or an antigen-binding fragment thereof is formulated in as sustained release formulation. In yet another embodiment, antibodies or fragments thereof which have higher avidity and/or higher affinity for one or more RSV antigens than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) and which comprises an Fc domain with increased affinity for the FcRn receptor relative to the Fc domain of SYNAGIS(copyright) are formulated in sustained release formulations.
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection in a mammal, preferably a human, comprising administering to said mammal a first dose of a prophylactically or therapeutically effective amount of one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens in a sustained release formulation, wherein said effective amount is a dose of 15 mg/kg or less of said antibodies or fragments thereof, which dose, preferably results in a serum titer of at least 2 xcexcg/ml (preferably at least 5 xcexcg/ml, at least 10 xcexcg/ml, at least 20 xcexcg/ml, at least 30 xcexcg/ml, or at least 40 xcexcg/ml) for at least 20 days (preferably at least 25, 30, 35 or 40 days) after the administration of the first dose and prior to the administration of a subsequent dose.
In one embodiment, a mammal, preferably a human, is administered a first dose of a prophylactically or therapeutically effective amount of SYNAGIS(copyright) or an antigen-binding fragment thereof in a sustained release formulation, wherein said effective amount is a dose of approximately 15 mg/kg of SYNAGIS(copyright) or an antigen-binding fragment thereof which dose results in a serum titer of at least 20 xcexcg/ml (preferably at least 30 xcexcg/ml, more preferably at least 40 xcexcg/ml, and most preferably at least 50 xcexcg/ml) for at least 30 days (preferably at least 35 days, more preferably at least 40 days, and most preferably at least 45 days) after the administration of the first dose and prior to the administration of a subsequent dose. In a preferred embodiment, a mammal, preferably a human, is administered a first dose of a prophylactically or therapeutically effective amount of SYNAGIS(copyright) or an antigen-binding fragment thereof in a sustained release formulation, wherein said effective amount is a dose of 15 mg/kg or less of SYNAGIS(copyright) or an antigen-binding fragment thereof which dose results in a serum titer of 20 xcexcg/ml (preferably at least 30 xcexcg/ml, more preferably at least 40 xcexcg/ml, and most preferably at least 50 xcexcg/ml) at least 30 days (preferably at least 35 days, more preferably at least 40 days, and most preferably at least 45 days) after the administration of the first dose and prior to the administration of a subsequent dose.
In another embodiment, a mammal, preferably a human, is administered a first dose of a prophylactically or therapeutically effective amount of one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) in a sustained release formulation, wherein said effective amount is a dose of less than 15 mg/kg (preferably 5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of said antibodies or antibody fragments which dose results in a serum titer of less than 30 xcexcg/ml (which is preferably at least 2 xcexcg/ml, more preferably at least 4 xcexcg/ml, and most preferably at least 6 xcexcg/ml) for at least 20 days (preferably at least 25, at least 30, at least 35, or at least 40 days) after the administration of the first dose and prior to the administration of a subsequent dose. In a preferred embodiment, a mammal, preferably a human, is administered a first dose of a prophylactically or therapeutically effective amount of one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) in a sustained release formulation, wherein said effective amount is a dose of less than 15 mg/kg of said antibodies or antibody fragments which dose results in a serum titer of 10 xcexcg/ml for at least 20 days (preferably at least 25, at least 30, at least 35 or at least 40 days) after the administration of the first dose and prior to the administration of a subsequent dose. In accordance with this embodiment, the prophylactically or therapeutically effective amount of the dose of the antibodies or antibody fragments is approximately 0.5 mg/kg, preferably 1 mg/kg, 1.5 mg/kg, 3 mg/kg, 5 mg/kg, 7.5 mg/kg, 10 mg/kg, 12 mg/kg, or 14 mg/kg. In another preferred embodiment, a mammal, preferably a human, is administered a first dose of a prophylactically or therapeutically effective amount of one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with higher avidity and/or higher affinity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) in a sustained release formulation, wherein said effective amount is a dose of 1.5 mg/kg of said antibodies or antibody fragments which dose results in a serum titer of 10 xcexcg/ml for at least 20 days (preferably at least 25, at least 30, at least 35, or at least 40 days) after the administration of the first dose and prior to the administration of a subsequent dose.
Additionally, the present invention provides sustained release compositions comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens, which sustained release compositions maintain a certain serum titer in a subject for a certain period of time without exceeding a particular serum titer. In one embodiment, a sustained release formulation comprising SYNAGIS(copyright) or an antigen-binding fragment thereof maintains a serum titer in a mammal, preferably a human, of approximately 25 xcexcg/ml (preferably 30 xcexcg/ml, more preferably 40 xcexcg/ml, and most preferably 50 xcexcg/ml) without exceeding a serum titer of approximately 100 xcexcg/ml (preferably 75 xcexcg/ml) for at least 20 days (preferably at least 25, 30, 35, or 40 days). In another embodiment, a sustained release formulation comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with a higher avidity and/or a higher affinity than previously known antibodies such as, e.g., SYNAGIS(copyright), maintains a serum titer in a mammal, preferably a human, of approximately 2 xcexcg/ml (preferably 6 xcexcg/ml, 10 xcexcg/ml, 20 xcexcg/ml, or 30 xcexcg/ml) without exceeding a serum titer of approximately 40 xcexcg/ml (preferably 75 xcexcg/ml) for at least 20 days (preferably at least 25, 30, 35, or 40 days).
The present invention encompasses methods of preventing, treating or ameliorating one or more symptoms of RSV infection in a mammal, preferably a human, by administering sustained release formulations of one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens and which have increased in vivo half-lives. In one embodiment, a sustained release formulation comprising HL-SYNAGIS or an antigen-binding fragment thereof is administered to a mammal, preferably a human, to prevent, treat, or ameliorate one or more symptoms associated with a RSV infection. In another embodiment, a sustained release formulation comprising one or more antibodies or fragments thereof which have higher avidity and/or higher affinity for one or more RSV antigens than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) and which comprises an Fc domain with increased affinity for the FcRn receptor relative to the Fc domain of SYNAGIS(copyright) are administered to a mammal, preferably a human, to prevent, treat, or ameliorate one or more symptoms associated with a RSV infection.
The present invention also provides pulmonary delivery systems for administering one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens. In particular, the present invention provides compositions for pulmonary delivery, said compositions comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens. SYNAGIS(copyright) or an antigen-binding fragment thereof can be incorporated into compositions for pulmonary delivery. HL-SYNAGIS or an antigen-binding fragment thereof can be incorporated into compositions for pulmonary delivery. One or more antibodies or fragments thereof that bind to one or more RSV antigens with higher affinity and/or higher avidity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) can be incorporated into compositions for pulmonary delivery. Further, one or more antibodies or fragments thereof which bind to one or more RSV antigens with higher affinity and/or higher avidity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments with an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for a RSV antigen) and which comprise an Fc domain with increased affinity for the FcRn receptor relative to the Fc domain of SYNAGIS(copyright) can be incorporated into compositions for pulmonary delivery.
The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection, said methods comprising administering to a mammal, preferably a human, a composition for pulmonary delivery comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens. In particular, the present invention provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection, said methods comprising administering to a mammal, preferably a human, a composition for pulmonary delivery comprising SYNAGIS(copyright) or fragments thereof. The present invention also provides methods for preventing, treating or ameliorating one or more symptoms associated with a RSV infection, said methods comprising administering to a mammal, preferably a human, a composition for pulmonary delivery comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with higher affinity and/or higher avidity than known antibodies such as, e.g., SYNAGIS(copyright) (e.g., antibodies or antibody fragments having an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for one or more RSV antigens).
In one embodiment, a first dose of a prophylactically or therapeutically effective amount of a composition comprising SYNAGIS(copyright) or an antigen-binding fragment thereof is administered to the lungs of a mammal, preferably a human, and results in an antibody concentration of at least 20 ng per mg of lung protein (preferably at least 40 ng/mg, at least 60 ng/mg, at least 80 ng/mg, at least 50 ng/mg, at least 75 ng/mg, at least 100 ng/mg, or at least 150 ng/mg) at least 20 days (preferably at least 25, 30, 35 or 40 days) after the administration of the first dose and prior to the administration of a subsequent dose. Preferably, the prophylactically or therapeutically effective amount is a dose of approximately 0.01 mg/kg, (preferably at least 0.1 mg/kg, at least 1 mg/kg, at least 2 mg/kg, at least 4 mg/kg, at least 5 mg/kg or at least 10 mg/kg) of SYNAGIS(copyright) or an antigen-binding fragment thereof.
In another embodiment, a first dose of a prophylactically or therapeutically effective amount of a composition comprising one or more antibodies or fragments thereof which immunospecifically bind to one or more RSV antigens with higher affinity and/or higher avidity than known antibodies such as, e.g., SYNAGIS(copyright), (e.g., antibodies or antibody fragments having an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for one or more RSV antigens) is administered to the lungs of a mammal, preferably a human and results in an antibody concentration of 20 ng per mg of lung protein (preferably at least 40 ng/mg, at least 60 ng/mg, at least 80 ng/mg, at least 50 ng/mg, at least 75 ng/mg, at least 100 ng/mg, or at least 150 ng/mg), at least 200 ng/mg, at least 250 ng/mg, at least 500 ng/mg, at least 750 ng/mg, at least 1 xcexcg/mg, at least 2 xcexcg/mg, at least 5 xcexcg/mg, at least 10 xcexcg/mg, at least 15 xcexcg/mg, or at least 25 xcexcg/mg) at least 20 days (preferably at least 25, 30, 35 or 40 days) at least 20 days (preferably at least 25, at least 30, at least 35 or at least 40 days after the administration of the first dose and prior to the administration of a subsequent dose. Preferably, the prophylactically effective amount is a dose of approximately 0.001 mg/kg, (preferably at least 0.005 mg/kg, at least 0.01 mg/kg, at least 0.05 mg/kg, at least 0.1 mg/kg, at least 1 mg/kg, at least 2 mg/kg, at least 4 mg/kg, at least 5 mg/kg or at least 10 mg/kg) of said antibodies or antibody fragments.
The present invention further provides detectable or diagnostic compositions comprising using antibodies or fragments thereof that immunospecifically bind to a RSV antigen, and methods for detecting or diagnosing a RSV infection utilizing said compositions.
The term xe2x80x9canalogxe2x80x9d as used herein refers to a polypeptide that possesses a similar or identical function as a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment but does not necessarily comprise a similar or identical amino acid sequence of a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment, or possess a similar or identical structure of a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment. A polypeptide that has a similar amino acid sequence refers to a polypeptide that satisfies at least one of the following: (a) a polypeptide having an amino acid sequence that is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the amino acid sequence of a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment described herein; (b) a polypeptide encoded by a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence encoding a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment described herein of at least 5 amino acid residues, at least 10 amino acid residues, at least 15 amino acid residues, at least 20 amino acid residues, at least 25 amino acid residues, at least 40 amino acid residues, at least 50 amino acid residues, at least 60 amino residues, at least 70 amino acid residues, at least 80 amino acid residues, at least 90 amino acid residues, at least 100 amino acid residues, at least 125 amino acid residues, or at least 150 amino acid residues; and (c) a polypeptide encoded by a nucleotide sequence that is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to the nucleotide sequence encoding a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment described herein. A polypeptide with similar structure to a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment described herein refers to a polypeptide that has a similar secondary, tertiary or quaternary structure of a RSV polypeptide, a fragment of a RSV, an antibody, or antibody fragment described herein. The structure of a polypeptide can determined by methods known to those skilled in the art, including but not limited to, X-ray crystallography, nuclear magnetic resonance, and crystallographic electron microscopy.
The term xe2x80x9cderivativexe2x80x9d as used herein refers to a polypeptide that comprises an amino acid sequence of a RSV polypeptide, a fragment of a RSV polypeptide, an antibody that immunospecifically binds to a RSV polypeptide, or an antibody fragment that immunospecifically binds to a RSV polypeptide which has been altered by the introduction of amino acid residue substitutions, deletions or additions. The term xe2x80x9cderivativexe2x80x9d as used herein also refers to a RSV polypeptide, a fragment of a RSV polypeptide, an antibody that immunospecifically binds to a RSV polypeptide, or an antibody fragment that immunospecifically binds to a RSV polypeptide which has been modified, i.e, by the covalent attachment of any type of molecule to the polypeptide. For example, but not by way of limitation, a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment may be modified, e.g., by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. A derivative of a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment may be modified by chemical modifications using techniques known to those of skill in the art, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Further, a derivative of a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment may contain one or more non-classical amino acids. A polypeptide derivative possesses a similar or identical function as a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, or antibody fragment described herein.
The term xe2x80x9ceffective neutralizing titerxe2x80x9d as used herein refers to the amount of antibody which corresponds to the amount present in the serum of animals (human or cotton rat) that has been shown to be either clinically efficacious (in humans) or to reduce virus by 99% in, for example, cotton rats. The 99% reduction is defined by a specific challenge of, e.g., 103 pfu, 104 pfu, 105 pfu, 106 pfu, 107 pfu, 108 pfu, or 109 pfu) of RSV.
The term xe2x80x9cepitopesxe2x80x9d as used herein refers to portions of a RSV polypeptide having antigenic or immunogenic activity in an animal, preferably a mammal, and most preferably in a human. An epitope having immunogenic activity is a portion of a RSV polypeptide that elicits an antibody response in an animal. An eptiope having antigenic activity is a portion of a RSV polypeptide to which an antibody immunospecifically binds as determined by any method well known in the art, for example, by the immunoassays described herein. Antigenic epitopes need not necessarily be immunogenic.
The term xe2x80x9cfragmentxe2x80x9d as used herein refers to a peptide or polypeptide comprising an amino acid sequence of at least 5 contiguous amino acid residues, at least 10 contiguous amino acid residues, at least 15 contiguous amino acid residues, at least 20 contiguous amino acid residues, at least 25 contiguous amino acid residues, at least 40 contiguous amino acid residues, at least 50 contiguous amino acid residues, at least 60 contiguous amino residues, at least 70 contiguous amino acid residues, at least contiguous 80 amino acid residues, at least contiguous 90 amino acid residues, at least contiguous 100 amino acid residues, at least contiguous 125 amino acid residues, at least 150 contiguous amino acid residues, at least contiguous 175 amino acid residues, at least contiguous 200 amino acid residues, or at least contiguous 250 amino acid residues of the amino acid sequence of a RSV polypeptide or an antibody that immunospecifically binds to a RSV polypeptide.
The term xe2x80x9chuman infantxe2x80x9d as used herein refers to a human less than 24 months, preferably less than 16 months, less than 12 months, less than 6 months, less than 3 months, less than 2 months, or less than 1 month of age.
The term xe2x80x9chuman infant born prematurelyxe2x80x9d as used herein refers to a human born at less than 40 weeks gestational age, preferably less than 35 weeks gestational age, who is less than 6 months old, preferably less than 3 months old, more preferably less than 2 months old, and most preferably less than 1 month old.
An xe2x80x9cisolatedxe2x80x9d or xe2x80x9cpurifiedxe2x80x9d antibody or fragment thereof is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the protein is derived, or substantially free of chemical precursors or other chemicals when chemically synthesized. The language xe2x80x9csubstantially free of cellular materialxe2x80x9d includes preparations of an antibody or antibody fragment in which the antibody or antibody fragment is separated from cellular components of the cells from which it is isolated or recombinantly produced. Thus, an antibody or antibody fragment that is substantially free of cellular material includes preparations of antibody or antibody fragment having less than about 30%, 20%, 10%, or 5% (by dry weight) of heterologous protein (also referred to herein as a xe2x80x9ccontaminating proteinxe2x80x9d). When the antibody or antibody fragment is recombinantly produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, 10%, or 5% of the volume of the protein preparation. When the antibody or antibody fragment is produced by chemical synthesis, it is preferably substantially free of chemical precursors or other chemicals, i.e., it is separated from chemical precursors or other chemicals which are involved in the synthesis of the protein. Accordingly such preparations of the antibody or antibody fragment have less than about 30%, 20%, 10%, 5% (by dry weight) of chemical precursors or compounds other than the antibody or antibody fragment of interest. In a preferred embodiment, antibodies of the invention or fragments thereof are isolated or purified.
An xe2x80x9cisolatedxe2x80x9d nucleic acid molecule is one which is separated from other nucleic acid molecules which are present in the natural source of the nucleic acid molecule. Moreover, an xe2x80x9cisolatedxe2x80x9d nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. In a preferred embodiment, nucleic acid molecules encoding antibodies of the invention or fragments thereof are isolated or purified.
The term xe2x80x9cfusion proteinxe2x80x9d as used herein refers to a polypeptide that comprises an amino acid sequence of an antibody or fragment thereof and an amino acid sequence of a heterologous polypeptide (e.g., a non-anti-RSV antigen antibody).
The term xe2x80x9chigh potencyxe2x80x9d as used herein refers to antibodies or fragments thereof that exhibit high potency as determined in various assays for biological activity (e.g., neutralization of RSV) such as those described herein. For example, high potency antibodies of the present invention or fragments thereof have an EC50 value less than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM as measured by a microneutralization assay described herein. Further, high potency antibodies of the present invention or fragments thereof result in at least a 75%, preferably at least a 95% and more preferably a 99% lower RSV titer in a cotton rat 5 days after challenge with 105 pfu relative to a cotton rat not administered said antibodies or antibody fragments. In certain embodiments of the invention, high potency antibodies of the present invention or fragments thereof exhibit a high affinity and/or high avidity for one or more RSV antigens (e.g., antibodies or antibody fragments having an affinity of at least 2xc3x97108 Mxe2x88x921, at least 2.5xc3x97108 Mxe2x88x921, at least 5xc3x97108 Mxe2x88x921, at least 109 Mxe2x88x921, at least 5xc3x97109 Mxe2x88x921, at least 1010 Mxe2x88x921, at least 5xc3x971010 Mxe2x88x921, at least 1011 Mxe2x88x921, at least 5xc3x971011 Mxe2x88x921, at least 1012 Mxe2x88x921, or at least 5xc3x971012 Mxe2x88x921 for one or more RSV antigens).
The term xe2x80x9chostxe2x80x9d as used herein refers to a mammal, preferably a human.
The term xe2x80x9chost cellxe2x80x9d as used herein refers to the particular subject cell transfected with a nucleic acid molecule and the progeny or potential progeny of such a cell. Progeny of such a cell may not be identical to the parent cell transfected with the nucleic acid molecule due to mutations or environmental influences that may occur in succeeding generations or integration of the nucleic acid molecule into the host cell genome.
In certain embodiments of the invention, a xe2x80x9cprophylactically effective serum titerxe2x80x9d is the serum titer in a mammal, preferably a human, that reduces the incidence of a RSV infection in said mammal. Preferably, the prophylactically effective serum titer reduces the incidence of RSV infections in humans with the greatest probability of complications resulting from RSV infection (e.g., a human with cystic fibrosis, bronchopulmonary dysplasia, congenital heart disease, congenital immunodeficiency or acquired immunodeficiency, a human who has had a bone marrow transplant, a human infant, or an elderly human). In certain other embodiments of the invention, a xe2x80x9cprophylactically effective serum titerxe2x80x9d is the serum titer in a cotton rat that results in a RSV titer 5 days after challenge with 105 pfu that is 99% lower than the RSV titer 5 days after challenge with 105 pfu of RSV in a cotton rat not administered an antibody or antibody fragment that immunospecifically binds to a RSV antigen.
In certain embodiments of the invention, a xe2x80x9ctherapeutically effective serum titerxe2x80x9d is the serum titer in a mammal, preferably a human, that reduces the severity, the duration and/or the symptoms associated with a RSV infection in said mammal. Preferably, the therapeutically effective serum titer reduces the severity, the duration and/or the number symptoms associated with RSV infections in humans with the greatest probability of complications resulting from a RSV infection (e.g., a human with cystic fibrosis, bronchopulmonary dysplasia, congenital heart disease, congenital immunodeficiency or acquired immunodeficiency, a human who has had a bone marrow transplant, a human infant, or an elderly human). In certain other embodiments of the invention, a xe2x80x9ctherapeutically effective serum titerxe2x80x9d is the serum titer in a cotton rat that results in a RSV titer 5 days after challenge with 105 pfu that is 99% lower than the RSV titer 5 days after challenge with 105 pfu of RSV in a cotton rat not administered an antibody or antibody fragment that immunospecifically binds to a RSV antigen.
As used herein, xe2x80x9cHL-SYNAGISxe2x80x9d is SYNAGIS(copyright) with one or more modifications in amino acid residues identified to be involved in the interaction between the Fc domain of SYNAGIS(copyright) and the FcRn receptor which results in an increase in the in vivo half-life of SYNAGIS(copyright) to greater than 21 days. An antigen-binding fragment of HL-SYNAGIS is a fragment of SYNAGIS(copyright) which immunospecifically binds to RSV F glycoprotein and has one or more modifications in amino acid residues identified to be involved in the interaction between the Fc domain of SYNAGIS(copyright) and the FcRn receptor, wherein said modifications result in an increase in the in vivo half-life of the antigen-binding fragment. In accordance with the invention, HL-SYNAGIS or an antigen-binding fragment thereof has an in vivo half-life of at least 25 days, preferably at least 30 days, more preferably at least 35 days, and most preferably at least 40 days.
The term xe2x80x9cRSV antigenxe2x80x9d refers to a RSV polypeptide or fragment thereof to which an antibody or antibody fragment immunospecifically binds. An RSV antigen also refers to an analog or derivative of a RSV polypeptide or fragment thereof to which an antibody or antibody fragment immunospecifically binds.
The term xe2x80x9cserum titerxe2x80x9d as used herein refers to an average serum titer in a population of least 10, preferably at least 20, and most preferably at least 40 subjects.
The term xe2x80x9cantibodies or fragments that immunospecifically bind to a RSV antigenxe2x80x9d as used herein refers to antibodies or fragments thereof that specifically bind to a RSV polypeptide or a fragment of a RSV polypeptide and do not non-specifically bind to other polypeptides. Antibodies or fragments that immunospecifically bind to a RSV polypeptide or fragment thereof may have cross-reactivity with other antigens. Preferably, antibodies or fragments that immunospecifically bind to a RSV polypeptide or fragment thereof do not cross-react with other antigens. Antibodies or fragments that immunospecifically bind to a RSV polypeptide can be identified, for example, by immunoassays or other techniques known to those of skill in the art.
To determine the percent identity of two amino acid sequences or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=number of identical overlapping positions/total number of positionsxc3x97100%). In one embodiment, the two sequences are the same length.
The determination of percent identity between two sequences can also be accomplished using a mathematical algorithm. A preferred, non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. U.S.A. 87:2264-2268, modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. U.S.A. 90:5873-5877. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecules of the present invention. BLAST protein searches can be performed with the XBLAST program parameters set, e.g., to scorexe2x88x9250, wordlength=3 to obtain amino acid sequences homologous to a protein molecule of the present invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402. Alternatively, PSI-BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI-Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., http://www.ncbi.nlm.nih.gov). Another preferred, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.
The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.